U.S. patent number 10,709,765 [Application Number 16/315,607] was granted by the patent office on 2020-07-14 for composition for inhibiting survival or growth of tubercle bacillus by adjusting polarization of macrophage, and method using same.
This patent grant is currently assigned to THE INDUSTRY & ACADEMIC COOPERATION IN CHUNGNAM NATIONAL UNIVERSITY (IAC). The grantee listed for this patent is THE INDUSTRY & ACADEMIC COOPERATION IN CHUNGNAM NATIONAL UNIVERSITY (IAC). Invention is credited to Chang-Hwa Song.
United States Patent |
10,709,765 |
Song |
July 14, 2020 |
Composition for inhibiting survival or growth of tubercle bacillus
by adjusting polarization of macrophage, and method using same
Abstract
The present invention provides a composition for inhibiting the
survival or growth of a tubercle bacillus, containing a material
which induces the polarization of a macrophage into an M1
macrophage, and a method for inhibiting the survival or growth of
the tubercle bacillus by using the composition.
Inventors: |
Song; Chang-Hwa (Daejeon,
KR) |
Applicant: |
Name |
City |
State |
Country |
Type |
THE INDUSTRY & ACADEMIC COOPERATION IN CHUNGNAM NATIONAL
UNIVERSITY (IAC) |
Daejeon |
N/A |
KR |
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Assignee: |
THE INDUSTRY & ACADEMIC
COOPERATION IN CHUNGNAM NATIONAL UNIVERSITY (IAC) (Daejeon,
KR)
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Family
ID: |
60912972 |
Appl.
No.: |
16/315,607 |
Filed: |
July 4, 2017 |
PCT
Filed: |
July 04, 2017 |
PCT No.: |
PCT/KR2017/007061 |
371(c)(1),(2),(4) Date: |
January 04, 2019 |
PCT
Pub. No.: |
WO2018/008932 |
PCT
Pub. Date: |
January 11, 2018 |
Prior Publication Data
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Document
Identifier |
Publication Date |
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US 20190307850 A1 |
Oct 10, 2019 |
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Foreign Application Priority Data
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Jul 5, 2016 [KR] |
|
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10-2016-0084770 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K
31/496 (20130101); A61K 31/133 (20130101); A61K
35/15 (20130101); A61P 31/06 (20180101); A61K
31/739 (20130101); A61K 31/4965 (20130101); A61K
45/06 (20130101); A61K 38/21 (20130101); A61K
38/217 (20130101); A61K 31/4409 (20130101); A61K
39/00 (20130101) |
Current International
Class: |
A61K
39/04 (20060101); A61K 38/21 (20060101); A61K
39/00 (20060101); A61P 31/06 (20060101); A61K
45/06 (20060101); A61K 35/15 (20150101); A61K
31/739 (20060101); A61K 31/133 (20060101); A61K
31/4409 (20060101); A61K 31/496 (20060101); A61K
31/4965 (20060101) |
Field of
Search: |
;424/9.1,9.2,234.1,248.1 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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100972304 |
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Jul 2010 |
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KR |
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1020130091061 |
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Aug 2013 |
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KR |
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Other References
Huang, Z., et al., PLOS ONE, vol. 10, pp. 1-16, Jun. 2015. cited by
examiner .
International Search Report for PCT/KR2017/007061 dated Aug. 30,
2017. cited by applicant .
Jae Seuk Park et al., "The Effect of IFN-y on the Phagocytosis of
Mycobacterium tuberculosis and Activation of Human Pulmonary
Alveolar Macrophage", Tuberculosis and Respiratory Diseases, Feb.
1998, pp. 36-44, vol. 45, No. 1. cited by applicant .
Simeone Marino et al., "Macrophage Polarization Drives Granuloma
Outcome during Mycobacterium tuberculosis Infection", Infection and
Immunity, Jan. 2015, pp. 324-338, vol. 83, No. 1. cited by
applicant .
Zikun Huang et al., "Mycobacterium tuberculosis-Induced
Polarization of Human Macrophage Orchestrates the Formation and
Development of Tuberculous Granulomas In Vitro", PLOS ONE, Jun. 19,
2015, pp. 1-16. cited by applicant.
|
Primary Examiner: Swartz; Rodney P
Claims
What is claimed is:
1. A composition for inhibiting the survival or proliferation of
Mycobacterium tuberculosis, the composition comprising: a material
inducing the polarization of macrophages into M1 macrophages, and
at least one anti-tuberculosis drug selected from the group
consisting of rifampicin, isoniazid, ethambutol, pyrazinamide, and
analogues thereof.
2. The composition of claim 1, wherein the material inducing
polarization of macrophages into M1 macrophages comprises at least
one selected from the group consisting of lipopolysaccharide,
interferon-gamma, IL-6, and TNF-.alpha..
3. The composition of claim 1, wherein the material inducing the
polarization of macrophages into M1 macrophages includes 1-20 ng/ml
of lipopolysaccharide and 1-20 ng/ml of interferon-gamma.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a U.S. National Stage of International Patent
Application No. PCT/KR2017/007061 filed Jul. 4, 2017, which claims
the priority benefit of Korean Patent Application No.
10-2016-0084770, filed on Jul. 5, 2016, in the Korean Intellectual
Property Office, the disclosures of which are incorporated herein
by reference.
TECHNICAL FIELD
The present invention relates to a method for inhibiting the
survival or proliferation of Mycobacterium tuberculosis, and a
method for adjusting the polarization of macrophages therefor.
BACKGROUND ART
Tuberculosis is a chronic infectious disease caused by infection
with Mycobacterium tuberculosis or other Mycobacterium species.
Tuberculosis is one of major diseases in developing countries, as
well as an increasing problem in developed countries, with about 8
million new patients and 3 million deaths each year. The infection
may be asymptomatic for a considerable period of time. However,
tuberculosis is most commonly manifested as an acute inflammation
of the lungs, resulting in fever and non-productive cough.
Moreover, if not treated, serious complications and death are
typically caused.
Tuberculosis can be treated using antibiotic therapy over a long
period of time, but such treatment is not sufficient to prevent the
spread of tuberculosis. Infected individuals may be asymptomatic,
but contagious, for a certain period of time. In addition, patient
behavior is difficult to monitor although the treatment regimen is
strictly followed. Some patients fail to complete the course of
treatment, which may result in ineffective treatment and the
development of drug resistance. Even after completion of the whole
course of treatment, infection with Mycobacterium tuberculosis
cannot be rooted out from infected individuals but still remains a
latent infection that can be reactivated.
In order to control the spread of tuberculosis, effective
vaccination and accurate early diagnosis of the disease are most
important. Currently, vaccination with live bacteria is the most
efficient way for inducing protective immunity. The most common
Mycobacterium used for this purpose is Bacillus Calmette-Guerin
(BCG), an avirulent strain of M. bovis. However, the stability and
efficacy of BCG is an issue of controversy, and some countries,
such as the US, do not vaccinate the general public with this
agent. Thus, studies continue on a technique for preventing
tuberculosis infection that is excellent in stability and efficacy,
compared with previous techniques.
Diagnosis of tuberculosis is typically achieved using a skin test,
which involves intradermal exposure to tuberculin purified protein
derivative (PPD). Antigen-specific T cell responses result in
measurable induration at the injection site by 48-72 hours after
injection, which indicates exposure to Mycobacterium antigens.
However, a problem of sensitivity and specificity arises with
regard to this test, and individuals vaccinated with BCG cannot be
distinguished from infected individuals.
Macrophages play an important role not only in natural immunity or
inflammatory responses but also in acquired immune responses, and
the representative function is presenting antigens to T cells
(antigen presentation). Macrophages receive protein antigens by
phagocytosis, decompose them into small peptides (antigen
processing), and bind the decomposed peptides to their MHCs, to
promote the activation of T cells again on the cell surface. Also,
macrophages act as the principal effector cells that indicate
immunological action in cell-mediated immune responses and humoral
immune responses, as well as having the function of suggesting
antigens required to initiate immune responses. Macrophages
activated by T cells perform the function of removing antigens from
a delayed-type hypersensitivity reaction, and macrophages have the
function of removing antigens bound by antibodies, with
phagocytosis.
The inventors of the present invention confirmed that Mycobacterium
tuberculosis disturbs the M1 polarization of macrophages by
secreting pathogenic factors and confirmed therefrom the inhibitory
efficacy of survival or proliferation of Mycobacterium tuberculosis
by inducing polarization of macrophages, thereby completing the
present invention.
DETAILED DESCRIPTION OF INVENTION
Technical Task
The present invention is to provide a composition for inhibiting
the survival or proliferation of Mycobacterium tuberculosis which
has different mechanism from previous drugs and causes relatively
fewer side effects, by using a material inducing the polarization
of macrophages, and a method for inhibiting the survival or
proliferation of Mycobacterium tuberculosis using the same.
Means for Solving the Task
An embodiment according to the present invention provides a
composition for inhibiting the survival or proliferation of
Mycobacterium tuberculosis, comprising a material inducing the
polarization of macrophages into M1 macrophages.
The material inducing polarization may comprise at least one
selected from the group consisting of lipopolysaccharide,
interferon-gamma and inflammatory cytokines, for example, IL-6 and
TNF-.alpha..
The composition may comprise 1-20 ng/ml of lipopolysaccharide and
1-20 ng/ml of interferon-gamma.
Also, the composition may further comprise at least one
anti-tuberculosis drug selected from the group consisting of
rifampicin, isoniazid, ethambutol, pyrazinamide, streptomycin or
analogues thereof.
The tuberculosis may be eye tuberculosis, skin tuberculosis,
adrenal tuberculosis, renal tuberculosis, epididymal tuberculosis,
lymph node tuberculosis, laryngeal tuberculosis, middle ear
tuberculosis, intestinal tuberculosis, multidrug-resistant
tuberculosis, pulmonary tuberculosis, sputum tuberculosis, bone
tuberculosis, throat tuberculosis, lymphatic gland tuberculosis,
lung deficiency, breast tuberculosis or spinal tuberculosis.
Another embodiment according to the present invention provides a
method for inhibiting the survival or proliferation of
Mycobacterium tuberculosis, comprising inducing the polarization of
macrophages into M1 macrophages using the composition.
The survival or proliferation of Mycobacterium tuberculosis may be
inhibited by inducing the polarization of macrophages into M1
macrophages by the composition, and thereafter inducing apoptosis
of the macrophages upon infection with Mycobacterium
tuberculosis.
Advantageous Effect
The present invention shows the effect of inhibiting Mycobacterium
tuberculosis by induction of macrophage polarization that was not
previously known. According to the present invention, it is
expected that not only typical Mycobacterium tuberculosis but also
multidrug-resistant Mycobacterium tuberculosis to existing drugs
can be prevented and treated.
BRIEF DESCRIPTION OF DRAWINGS
FIGS. 1A to 1D are electrophoresis photographs of protein
expression using western blotting, which shows the expression of
macrophage M1/M2 polarization markers upon infection with
Mycobacterium tuberculosis in macrophages.
FIGS. 2A and 2B are drawings which show apoptosis induced by
infection with Mycobacterium tuberculosis in M1/M2 macrophages.
FIG. 2C is an electrophoresis photograph of protein expression
using western blotting, which shows the activation of Caspase-3,
Bax and Bcl-2 in M1/M2 macrophages infected with Mycobacterium
tuberculosis.
FIG. 2D is an electrophoresis photograph of protein expression
using western blotting, which shows the intracellular transport of
cytochrome C after isolating M1/M2 macrophages infected with
Mycobacterium tuberculosis into cytoplasm and mitochondria.
FIGS. 3A and 3B are graphs which measure the number of
Mycobacterium tuberculosis intracellularly surviving in M1/M2
macrophages infected with Mycobacterium tuberculosis.
FIGS. 3C, 3D, 3E and 3F are drawings which show the intracellular
survival of Mycobacterium tuberculosis in M1/M2 macrophages
infected with Mycobacterium tuberculosis, through a fluorescence
microscope.
FIG. 4 is a graph which measures the number of intracellular
Mycobacterium tuberculosis in M1/M2 macrophages infected with
Mycobacterium tuberculosis upon the use of an anti-tuberculosis
drug in combination.
FIG. 5A is an electrophoresis photograph of protein expression
using western blotting, which shows the activation of CHOP and
Caspase-3 in lung tissues 20 days after infection with
Mycobacterium tuberculosis through the nasal cavity in M1/M2 mouse
models.
FIG. 5B is a graph which measures the number of Mycobacterium
tuberculosis surviving in the lung tissues of M1/M2 mouse models
infected with Mycobacterium tuberculosis.
FIG. 5C is a graph which measures the number of Mycobacterium
tuberculosis surviving in the lung tissues in M1/M2 mouse models
infected with Mycobacterium tuberculosis upon the use of an
anti-tuberculosis drug in combination.
BEST MODE FOR CARRYING OUT THE INVENTION
Interferon-gamma allows to have more enhanced antigen presenting
ability than macrophages before encountering antigens, and also to
increase complement-mediated phagocytosis and produce inflammatory
cytokines. Such macrophages are so-called classically activated
macrophages (M1).
Helper T lymphocytes can be divided into two major types, Th1 and
Th2 cells, based on secreted cytokines. Cytokines which are mainly
secreted by Th2 cells, such as IL-4, set macrophages in the state
opposite to the activation induced by interferon-gamma, for
example, increasing the expression of major histocompatibility
antigen class. Macrophages in such state are called alternatively
activated macrophages (M2).
In order to achieve the aforementioned objective, one embodiment
according to the present invention provides a composition for
inhibiting the survival or proliferation of Mycobacterium
tuberculosis, comprising a material inducing the polarization of
macrophages into M1 macrophages.
The material inducing polarization may comprise at least one
selected from the group consisting of lipopolysaccharide,
interferon-gamma and inflammatory cytokines, for example, IL-6 and
TNF-.alpha..
Specifically, the composition may comprise 1-20 ng/ml of
lipopolysaccharide and 1-20 ng/ml of interferon-gamma.
Also, the composition may further comprise at least one
anti-tuberculosis drug selected from the group consisting of
rifampicin, isoniazid, ethambutol, pyrazinamide or analogues
thereof.
The tuberculosis may be eye tuberculosis, skin tuberculosis,
adrenal tuberculosis, renal tuberculosis, epididymal tuberculosis,
lymph node tuberculosis, laryngeal tuberculosis, middle ear
tuberculosis, intestinal tuberculosis, multidrug-resistant
tuberculosis, pulmonary tuberculosis, sputum tuberculosis, bone
tuberculosis, throat tuberculosis, lymphatic gland tuberculosis,
lung deficiency, breast tuberculosis or spinal tuberculosis.
Another embodiment according to the present invention provides a
method for inhibiting the survival or proliferation of
Mycobacterium tuberculosis, comprising inducing the polarization of
macrophages into M1 macrophages using the composition.
The method may further comprise steps before and after infection
with Mycobacterium tuberculosis.
The survival or proliferation of Mycobacterium tuberculosis may be
inhibited by inducing the polarization of macrophages into M1
macrophages by the composition, and thereafter inducing apoptosis
of the macrophages upon infection with Mycobacterium
tuberculosis.
Hereinafter, preferred embodiments of the method for inhibiting the
survival and proliferation of intracellular Mycobacterium
tuberculosis using the polarization of macrophages according to the
present invention will be described in detail with reference to the
accompanying drawings. The present invention is not limited to the
embodiments described below but may be implemented into various
embodiments. The present embodiments are provided simply to
complete the disclosure of the present invention and help a person
skilled in the art completely understand the scope of the present
invention.
MODES FOR CARRYING OUT THE INVENTION
Example 1
In Vitro Experiment for Assessing Inhibitory Effect of Polarization
of Macrophage on Survival of Mycobacterium tuberculosis
1. Step of Confirming which Roles the Polarization of Macrophage
Plays Upon Infection with Mycobacterium tuberculosis
In order to analyze which roles the polarization of macrophages
play upon infection with Mycobacterium tuberculosis, macrophage
polarization induced by infection with Mycobacterium tuberculosis
was confirmed. That is, the expression level of polarization
markers was confirmed over time, after infection with pathogenic
Mycobacterium tuberculosis (Mycobacterium tuberculosis H37Rv) and
non-pathogenic Mycobacterium tuberculosis (M tuberculosis H37Ra) in
the rate of 1 bacterium per cell, using bone marrow-derived
macrophages (BMDMs), mouse-derived macrophages.
FIG. 1A is the result of confirmation on protein expression level
of phospho-STAT1 (p-STAT1), a marker indicating M1 macrophage
polarization, and phospho-STAT3 (p-STAT3) and phospho-STAT6
(p-STATE), markers indicating M2 macrophage polarization, in
macrophages infected with Mycobacterium tuberculosis, using a
western blotting method. It demonstrates that the expression of
phospho-STAT3 and phospho-STAT6 which indicate M2 macrophage
polarization increases over cultivation time in macrophages
infected with pathogenic Mycobacterium tuberculosis. In contrast,
it could be confirmed that the expression of phospho-STAT1 which
indicates M1 macrophage polarization increases in macrophages
infected with non-pathogenic Mycobacterium tuberculosis. (In the
drawings, STAT1, STAT3 and STATE are internal controls; the rest is
the same as above.)
FIGS. 1B, 1C and 1D are the result of confirmation on protein
expression level of iNOS, NICD, FPR2 and RhoA which indicate M1
macrophage polarization, and Arginase 1, KLF4 and Rac1 which
indicate M2 macrophage polarization, in macrophages infected with
H37Rv or H37Ra, using a western blotting method. It demonstrates
that the expression of Arginase 1, KLF4 and Rac1 which indicate M2
macrophage polarization increases over cultivation time in
macrophages infected with pathogenic Mycobacterium tuberculosis. In
contrast, it could be confirmed that the expression of iNOS, NICD,
FPR2 and RhoA which indicate M1 macrophage polarization increases
in macrophages infected with non-pathogenic Mycobacterium
tuberculosis. (In the drawings, .beta.-actin is an internal
control; the rest is the same as above.)
Also, Rv and Ra shown in FIG. 1 denote macrophages which are
infected with H37Rv (pathogenic Mycobacterium tuberculosis) or
H37Ra (non-pathogenic Mycobacterium tuberculosis) in the
experiment. It means that the polarization of macrophages varies
depending on the pathogenicity of Mycobacterium tuberculosis.
2. Step of Polarizing Mouse-Derived Macrophages into M1/M2
Mouse-derived macrophages were stimulated for 24 hours with
lipopolysaccharide (10 ng/ml) and interferon-gamma (IFN.gamma.) (10
ng/ml) to prepare M1 macrophages, and were stimulated for 24 hours
with IL-4 (10 ng/ml) and IL-13 (10 ng/ml) to prepare M2
macrophages.
3. Step of Confirming Induction of Apoptosis after Infecting the
Macrophages with Mycobacterium tuberculosis
FIG. 2A confirmed apoptosis using FACS after staining M1/M2
macrophages infected with Mycobacterium tuberculosis with
Annexin-V/PI. FIG. 2B is a drawing which indicates the percentage
of Annexin-V+/PI- cells by graph. It could be understood that upon
infection with Mycobacterium tuberculosis, apoptosis is induced in
M1 macrophages at a significantly higher rate than in M2
macrophages. FIG. 2C confirmed the induction of apoptosis in M1/M2
macrophages infected with Mycobacterium tuberculosis, using
Caspase-3, Bax and Bcl-2. Similar to the result above, the
expression of Caspase-3 and Bax increased, whereas the expression
of Bcl-2 decreased, in M1 macrophages.
4. Step of Confirming Intracellular Transport of Cytochrome C after
Infecting the Macrophages with Mycobacterium tuberculosis
FIG. 3D is the result of confirmation on the intracellular
transport of cytochrome C after isolating M1/M2 macrophages
infected with Mycobacterium tuberculosis into cytoplasm and
mitochondria. It was confirmed that upon infection with
Mycobacterium tuberculosis, cytochrome C transport increases from
mitochondria to cytoplasm in M1 macrophages. (In the drawings,
.alpha.-tubulin is a cytoplasm control, and COX IV is a
mitochondria control.)
According to the previous presentation of the present inventors, it
was reported that apoptosis induced by Mycobacterium tuberculosis
in macrophages is a significant mechanism for inhibition of
intracellular survival of Mycobacterium tuberculosis. Therefore, it
can be understood from the aforementioned experimental result that
upon infection with Mycobacterium tuberculosis, M1 macrophages
induce apoptosis and is thus more effective in inhibiting
proliferation of Mycobacterium tuberculosis.
Example 2
Case of Using a Material Adjusting Macrophage Polarization and an
Anti-Tuberculosis Drug in Combination
1. Experiment for Analyzing which Roles the Polarization of
Macrophage Plays Upon Infection with Mycobacterium tuberculosis
In order to analyze which roles the polarization of macrophages
play upon infection with Mycobacterium tuberculosis, the number of
Mycobacterium tuberculosis within M1/M2 macrophages was
measured.
FIGS. 3A and 3B are graphs which show the result of cultivation for
24 and 48 hours under complete medium conditions, after washing
extracellular Mycobacterium tuberculosis in M1/M2 macrophages which
were infected with pathogenic Mycobacterium tuberculosis and
non-pathogenic Mycobacterium tuberculosis, respectively, 3 hours
after infection. In order to determine the number of
intracellularly surviving Mycobacterium tuberculosis, cells were
cultured in 7H10 agar medium for 14 to 21 days to calculate the
total number of viable bacteria. FIGS. 3C and 3E are drawings which
show Mycobacterium tuberculosis surviving in M1/M2 macrophages 48
hours after infection, using a fluorescence microscope, which are
indicated by graph in FIGS. 3D and 3F. It could be confirmed that
upon infection with Mycobacterium tuberculosis, Mycobacterium
tuberculosis was controlled in M1 macrophages more effectively than
in M2 macrophages.
2. Experiment for Confirming the Inhibitory Effect on the Survival
of Mycobacterium tuberculosis by Using a Material Adjusting
Macrophage Polarization and an Anti-Tuberculosis Drug in
Combination
FIG. 4 measured the number of intracellular Mycobacterium
tuberculosis after washing extracellular Mycobacterium tuberculosis
and treating an anti-tuberculosis drug (Rifampicin: RIF,
Ethambutol: EMB, Isoniazid: INH, Pyrazinamide: PZA) or an
anti-tuberculosis drug together with a material inducing M1/M2
macrophage polarization (LPS+IFN.gamma. or IL-4+IL-13) in
combination thereon, 3 hours after infecting macrophages with
Mycobacterium tuberculosis, and performing cultivation for 24 and
48 hours. As a result of experiment, it was confirmed that upon
treatment of an anti-tuberculosis drug alone, the number of
Mycobacterium tuberculosis was reduced, and particularly, the
survival of Mycobacterium tuberculosis was inhibited more
efficiently in cells induced into M1 polarization. By comparison,
it was demonstrated that the survival of Mycobacterium tuberculosis
was not effectively inhibited in M2 macrophages, in spite of
treatment of an anti-tuberculosis drug.
Therefore, based on the aforementioned experimental result of M1
macrophages and the already known fact that an anti-tuberculosis
drug inhibits the survival of Mycobacterium tuberculosis, it was
determined that the adjustment of macrophage polarization together
with the administration of an anti-tuberculosis drug would be more
effective in killing Mycobacterium tuberculosis, and the hypothesis
was verified.
Example 3
In Vivo Experiment for Assessing Inhibitory Effect of Polarization
of Macrophage on Survival of Mycobacterium tuberculosis (Mouse
Model)
1. Step of Polarizing Mouse-Derived Macrophages into M1/M2
Mouse-derived macrophages were stimulated for 24 hours with
lipopolysaccharide (10 ng/ml) and interferon-gamma (IFN.gamma.) (10
ng/ml) to prepare M1 macrophages, and were stimulated for 24 hours
with IL-4 (10 ng/ml) and IL-13 (10 ng/ml) to prepare M2
macrophages.
2. Experiment for Assessing Inhibitory Effect of the Polarization
of Macrophages on Survival of Mycobacterium tuberculosis after
Infecting the Macrophages with Mycobacterium tuberculosis
FIG. 5A is the result of protein expression level of CHOP and
Caspase-3 in the lung tissues of M1/M2 mouse models 20 days after
infection by infecting M1/M2 mouse models with Mycobacterium
tuberculosis through the nasal cavity, using a western blotting
method. It could be observed that the activation of apoptosis is
remarkably higher in the lung tissues of M1 mouse model than in
those of M2 mouse model. FIG. 5B is a graph which shows the result
of measuring the number of Mycobacterium tuberculosis surviving in
the lung tissues of M1/M2 mouse models 3, 7 and 20 days after
infection with Mycobacterium tuberculosis. FIG. 5B is a graph which
shows the result of measuring the number of Mycobacterium
tuberculosis surviving in the lung tissues of M1/M2 mouse models 3
days after infection with Mycobacterium tuberculosis, after
infecting M1/M2 mouse models with Mycobacterium tuberculosis, and
then administering 10 mg/kg of an anti-tuberculosis drug (a mixture
of RIF, EMB, INH and PZA) through drinking water (1.5 ml/10 g mouse
weight). It could be confirmed that the survival of Mycobacterium
tuberculosis is more advantageous in the lung of M2 mouse model
than in that of mouse model, even upon administration of the
anti-tuberculosis drug. Similar to the result of in vitro
experiment, in in vivo experiment, it appears that it would be more
effective in inhibiting the proliferation of Mycobacterium
tuberculosis by inducing apoptosis in M1 mouse model upon infection
with Mycobacterium tuberculosis.
It was confirmed from the above experiment that the polarization of
macrophages is adjusted according to the pathogenicity of
Mycobacterium tuberculosis, and that upon infection with
Mycobacterium tuberculosis, macrophages polarized into M1 have a
remarkably high ability to inhibit the intracellular growth of
Mycobacterium tuberculosis using apoptosis. By comparison, M2
macrophages provide an advantageous environment to the
intracellular survival of Mycobacterium tuberculosis, and thus
Mycobacterium tuberculosis having stronger pathogenicity induces
the polarization into M2 macrophages. The present result proves
that it is possible to control the intracellular survival of
Mycobacterium tuberculosis by adjusting the polarization of
macrophages.
* * * * *